Substantially increased sensitivity of the spot-ELISA for the detection of anti-insulin antibody-secreting cells using a capture antibody and enzyme-conjugated insulin.
This paper describes an antibody capture spot-ELISA for the detection of anti-insulin antibody-secreting cells. The assay is based on the binding of secreted antibodies by immobilised isotype-specific capture antibodies and subsequent detection of insulin-specific antibodies with a conjugate of human insulin and alkaline phosphatase (HI-AP). Compared with the conventional approach, using antigen for coating and employing an enzyme-linked detecting antibody, this technique improved the detection of murine cells secreting anti-insulin antibodies of different IgG subclasses.
Vos
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Q
Department of Cell Biology, Immunology and Genetics, Erasmus University, Rotterdam, The Netherlands.
Claassen
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E
Benner
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R
D000818:Animals; D000921:Antibody-Producing Cells / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D007074:Immunoglobulin G / Q000032:analysis; D007330:Insulin Antibodies / Q000032:analysis*; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C
D007074:Immunoglobulin G; D007330:Insulin Antibodies